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Remarkable Improvement of Methylglyoxal Synthase Thermostability by His–His Interaction

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Abstract

Lately it has been proposed that interaction between two positively charged side chains can stabilize the folded state of proteins. To further explore this point, we studied the effect of histidine–histidine interactions on thermostability of methylglyoxal synthase from Thermus sp. GH5 (TMGS). The crystal structure of TMGS revealed that His23, Arg22, and Phe19 are in close distance and form a surface loop. Here, two modified enzymes were produced by site-directed mutagenesis (SDM); one of them, one histidine (TMGS-HHO), and another two histidines (TMGS-HHHO) were inserted between Arg22 and His23 (HO). In comparison with the wild type, TMGS-HHO thermostability increased remarkably, whereas TMGS-HHHO was very unstable. To explore the role of His23 in the observed phenomenon, the original His23 in TMGS-HHHO was replaced with Ala (TMGS-HHA). Our data showed that the half-life of TMGS-HHA decreased in relation to the wild type. However, its half-life increased in comparison with TMGS-HHHO. These results demonstrated that histidine–histidine interactions at position 23 in TMGS-HHO probably have the main role in TMGS thermostability.

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Abbreviations

TMGS:

Thermus sp. GH5 methylglyoxal synthase

HO :

TMGS original histidine

TMGS-HHO and TMGS-HHHO :

Mutant TMGS

DHAP:

Dihydroxyacetone phosphate

SDM:

Site-directed mutagenesis

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Acknowledgment

The authors express their gratitude to the research council of Tarbiat Modares University for the financial support during the course of this project.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box 14115-175, Tehran, Iran

    Malihe Mohammadi, Mona Atabakhshi Kashi, Manoochehr Mirshahi & Khosro Khajeh

  2. Department of Biological Sciences, Institute for Advanced Studies in Basic Sciences, Zanjan, Iran

    Shekufeh Zareian

Authors
  1. Malihe Mohammadi
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  2. Mona Atabakhshi Kashi
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  3. Shekufeh Zareian
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  4. Manoochehr Mirshahi
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  5. Khosro Khajeh
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Correspondence to Khosro Khajeh.

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Mohammadi, M., Kashi, M.A., Zareian, S. et al. Remarkable Improvement of Methylglyoxal Synthase Thermostability by His–His Interaction. Appl Biochem Biotechnol 172, 157–167 (2014). https://doi.org/10.1007/s12010-013-0404-y

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  • DOI: https://doi.org/10.1007/s12010-013-0404-y

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